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Application: Study enzyme kinetics and screen small molecule inhibitors for drug discovery and high throughput screening (HTS) applications.
Background: Cyclin-dependent kinase 9 (CDK9) is the catalytic subunit of the positive transcription elongation factor b (P-TEFb) complex, which phosphorylates the C-terminal domain of RNA polymerase II, a key player in the production of mature RNA. P-TEFb also includes cyclinT. CDK9 translocates to the nucleus once it gets auto phosphorylated, where it can associate with cyclinT. Hyperactivation of cyclins is known to result in cancer and the development of resistance to therapy, and CDK9-cyclinT is involved in the transcription of oncogenes. The inhibition or degradation of specific CDK-cyclin complexes could prove beneficial in cancer treatment, but the development of specific inhibitors has been difficult. Recently a small molecule degrader specific for CDK9-cyclinT was identified, LL-K9-3, which resulted in lower levels of androgen receptor (AR) and cMyc in 22RV1 cells. LL-K9-3 thus seemed more effective than SNS032, its parental CDK9 inhibitor, and the CDK9 PROTAC Thal-SNS032. Further research into new and more specific degrader molecules will prove beneficial for cancer therapy.
Contraindications: The final concentration of DMSO in the assay should not exceed 1%.
Description: The CDK9/CyclinT Kinase Assay Kit is designed to measure CDK9/CyclinT kinase activity for screening and profiling applications using Kinase-Glo™ Max as a detection reagent. The assay kit comes in a convenient 96-well or 384-well format, with enough purified recombinant CDK9/CyclinT kinase, kinase substrate, ATP, and kinase assay buffer for 100 enzyme reactions.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Supplied As: The CDK9 Assay Kit comes in a convenient 96-well format, with enough purified recombinant CDK9/CyclinT enzyme, CDK substrate peptide 2, ATP and kinase assay buffer for 100 enzyme reactions.
Uniprot: CDK9: P50750; CyclinT1: O60563
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Napolitano G, et al., 2003 J Cell Physiol. 197(1):1-7.Li J., et al., 2022 J Med Chem. 65(16):11034-11057.